Plasmídeos associados à nanopartículas biodegradáveis para melhorar as propriedades de estabilidade: uma aplicação biotecnológica

Abstract

Plasmids (pDNAs) are circular DNAs presented mainly in bacteria, with biotechnological importance in the area of gene therapy and DNA vaccines. However, even though they are safe and affordable, these vectors have restrictions related to low stability and low efficiency in cell transfection. Therefore, this work aims to elucidate physicochemical and biological stability parameters of plasmids pVAX1, pVAX1LacZ and nTSApVAX1 – such as size, temperature and blood nucleases’ concentration – and to develop new nanoformulations that could guarantee greater protection to the plasmids, through use of functionalized with PEI (NC) and white biodegradable polymer nanoparticles (NB). For the evaluation of plasmid stability and viability, 1% agarose gel electrophoresis was performed. The results showed that all pDNAs are more degradable in high concentrations of mouse and human serum. Regarding size, the larger plasmid, pVAX1LacZ, was more susceptible to nuclease action; meanwhile temperature directly influenced the stability of these plasmids. Concerning the nanoformulations, it was found that white nanoparticles were not efficient in protecting the plasmids, while PEI functionalized nanoparticles proved to be efficient in protecting pDNAs.Through this study, it was possible to obtain a promising biotechnological alternative, which gives greater stability to the plasmids, enabling future applications in DNA therapies.